Mouse anti-HPV-16 E7 antibody and rabbit anti‑human MAOA and MMP-2 antibodies were obtained from Abcam (Cambridge, UK). Mouse anti‑human Ki-67, rabbit anti‑human E‑cadherin, N-cadherin, Vimentin, Slug, Twist1, MMP-9, p-ERK1/2, ERK1/2, p-AKT, AKT, and β-actin monoclonal antibodies, and horseradish peroxidase‑conjugated secondary antibodies were from Cell Signaling Technology Inc. (Beverly, MA, USA). Mouse anti-human HIF-1α monoclonal antibody was purchased from BD Transduction Laboratories (San Diego, CA, USA). The RNA extraction kits (RNAprep Pure FFPE kits) were obtained from Tiangen Biotech Co., Ltd. (Beijing, China). The reverse transcription (RT) kit (PrimeScript™ RT reagent kits) and the qPCR analysis kit (SYBR Premix Ex Taq™ II) were purchased from Takara Biotechnology Co., Ltd. (Dalian, China). PD98059 was purchased from Cell Signaling Technology Inc. (Beverly, MA, USA). The human vascular endothelial growth factor (VEGF) ELISA kit was purchased from Boster Biological Technology Co.Ltd (Wuhan, China). The ROS assay kit was purchased from Beyotime Biotechnology Co. Ltd (Shanghai, China).
Mouse anti human ki 67
Manufactured by Cell Signaling Technology
Sourced in United States
Mouse anti-human Ki-67 is a monoclonal antibody that recognizes the Ki-67 protein, a cellular marker for proliferation. It is used to detect the presence and localization of the Ki-67 antigen in human tissues.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti human e cadherin, by Cell Signaling Technology (1 mentions)
Vegf elisa kit, by Boster Bio (1 mentions)
Twist1, by Cell Signaling Technology (1 mentions)
Mouse anti human hif 1α monoclonal antibody, by BD (1 mentions)
P erk1 2, by Cell Signaling Technology (1 mentions)
Pd98059, by Cell Signaling Technology (1 mentions)
Primescript rt reagent kit, by Takara Bio (1 mentions)
Mmp 9, by Cell Signaling Technology (1 mentions)
Sybr premix ex taq 2, by Takara Bio (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
N cadherin, by Cell Signaling Technology (1 mentions)
Vimentin, by Cell Signaling Technology (1 mentions)
Erk1 2, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Ros assay kit, by Beyotime (1 mentions)
Hrp conjugated secondary antibody, by ZSGB-BIO (1 mentions)
Cell culture dishes, by Corning (1 mentions)
Penicillin mixture, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
5 protocols using mouse anti human ki 67
1
Comprehensive Protein and Gene Expression Analysis in Cancer
2
Immunohistochemical Analysis of Tumor Markers
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Serial sections at 4 μm thickness were obtained from paraffin-embedded samples. Sections were incubated with rabbit anti-human UBN2 (1:100 dilution; BD Biosciences), mouse anti-human Ki-67 (1:200 dilution, Cell Signalling Technology, MA, USA) and mouse anti-human Cleaved Caspase-3 (1:200 dilution, Abcam, USA) overnight at 4°C after antigen retrieval and the block of non-specific binding. The sections were incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies (ZSGB, Beijing, China). Expression was visualized, and further independently scored based on the percentage of positively stained tumor cells by two observers.
3
Glioma Cell Line U251 Cultivation
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The glioma cell line U251 was purchased from ATCC, USA. Fetal bovine serum, high glucose DMEM, penicillin mixture, and trypsin were all purchased from Gibco, USA. LCA powder (molecular formula: C21H22O4; molecular weight: 338.40; purity ≥ 96.0%), dimethyl sulfoxide preparation, and methyl thiazolyl tetrazolium (MTT) kit were purchased from Sigma, USA. A 4% paraformaldehyde was purchased from Invitrogen, USA. Radio immunoprecipitation assay (RIPA) cell lysate was purchased from Beyotime Institute of Biotechnology, China. Bicinchoninic acid (BCA) kit and horseradish peroxidase-labeled goat anti-mouse IgG antibody were purchased from Thermo Company, USA. The ultrasensitive ECL chemiluminescence kit was purchased from Advansta, USA. Mouse anti-human Ki-67, β-catenin, GSK3β, Axin2, c-myc, Bcl-2, Bax, and GAPDH were all purchased from Cell Signaling, USA. Puromycin was purchased from Merck, USA. Lentiviral Vector Particle was purchased from China Jikai Gene. The cell culture dishes were purchased from Corning Costar, USA.
4
Immunohistochemical Scoring of Ran and Ki-67
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The slides were first incubated on a 60 °C heating panel for 1 h, then deparaffinised, subjected to antigen retrieval and endogenous peroxidase inactivation and incubated with the following primary antibodies: mouse anti-human Ran (BD Biosciences, CA, USA) and mouse anti-human Ki-67 (Cell Signalling Technology, MA, USA). Then the sections were incubated with horseradish peroxidase (HRP)-conjugated goat anti-rabbit or goat anti-mouse secondary antibodies (ZSGB, Beijing, China). The target molecules were detected in situ with diaminobenzidine (ZSGB), and the sections were counterstained with haematoxylin–eosin (ZSGB). Expression was visualised and classified based on the intensity of staining and the percentage of positive cells. The intensity score was divided into four grades: negative (0), weak (1), moderate (2), and strong (3). The percentage score was also divided into four grades: ≤1% (0), 2–25% (1), 26–50% (2), 51–75% (3), and ≥75% (4). The histological score was calculated by the following formula: histological score = intensity score × percentage score. An overall score of 0–12 was graded as negative (−, score: 0), weak (+, score: 1–4), moderate (++, score: 5–8) or strong (+++, score: 9–12).
5
Cell Cycle, Proliferation, and Apoptosis Analysis
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Flow cytometry was used to examine DAPI for cell cycle analysis, Ki-67 for cell proliferation, and cleaved caspase 3 expression for apoptosis following drug treatment. Cells were fixed with 4% formaldehyde and permeabilized using ice-cold methanol. The cells were then incubated with DAPI (10 mg/mL, 1:1000), mouse anti-human Ki-67 (Cell Signaling #9449, 1:400), and rabbit anti-human cleaved caspase 3 antibodies (Cell Signaling #9661, 1:800) at room temperature for 1 hour. Anti-mouse IgG DyLight 594 (Invitrogen #35511, 1:100) and anti-rabbit IgG DyLight 488 (Invitrogen #35553, 1:100) were used to detect anti-Ki-67 and anti-cleaved caspase 3 antibodies, respectively. The experiments were carried out on a CytoFLEX Flow Cytometry (Beckman Coulter, CA, USA), and the data was analyzed with FlowJo v10 from BD Biosciences (Franklin Lakes, NJ, USA).
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